金葡菌杀白细胞素S组分LukS-PV抑制急性髓系白血病细胞增殖的机制研究*

doi:10.3969/j.issn.1671-2587.2020.06.011

临床输血与检验 ›› 2020, Vol. 22 ›› Issue (6): 603-609.DOI: 10.3969/j.issn.1671-2587.2020.06.011

金葡菌杀白细胞素S组分LukS-PV抑制急性髓系白血病细胞增殖的机制研究^*

施岚, 许良飞, 聂正超, 常文娇, 马筱玲

230001 中国科学技术大学附属第一医院检验科

收稿日期:2020-07-28 出版日期:2020-12-20 发布日期:2021-01-18
作者简介:施岚(1989-),女,安徽阜阳人,检验技师,硕士,主要从事血液学研究,(E-mail)lanshi1008@126.com
基金资助:
*本课题受安徽省自然科学基金(No.1808085QH259)、中央高校基础研究基金(No.WK9110000107)资助

Study on the Inhibitory Mechanism of Staphylococcus Aureus Leukocidin S Component LukS-PV on the Proliferation in Acute Myeloid Leukemia Cells

SHI Lan, XU Liang-fei, NIE Zheng-chao, et al

Department of Laboratory,the First Affiliated Hospital of University of Science and Technology of China,Hefei 230001

Received:2020-07-28 Online:2020-12-20 Published:2021-01-18

摘要/Abstract

摘要： 目的探讨金葡菌杀白细胞素S组分LukS-PV通过下调甲基转移酶SET8对急性髓系白血病(AML)细胞THP-1和HL-60增殖的影响。方法采用实时荧光定量PCR(qRT-PCR)和Western blot检测不同浓度LukS-PV处理后THP-1和HL-60细胞的SET8 mRNA和蛋白的表达水平。将敲低SET8表达的慢病毒载体转染至THP-1和HL-60细胞中,qRT-PCR和Western blot检测SET8敲低效率,EdU实验检测敲低SET8对细胞增殖的影响。在敲低SET8的基础上,加入LukS-PV处理,通过EdU检测细胞增殖探究LukS-PV是否通过下调SET8抑制白血病细胞THP-1和HL-60的增殖。结果在AML细胞系THP-1和HL-60中,与PBS对照组相比,LukS-PV处理组SET8及其修饰位点H4K20me1的表达水平明显降低并具有浓度依赖性(P<0.05)。与阴性对照组相比,在THP-1和HL-60细胞转染敲低SET8慢病毒载体后,SET8表达水平明显降低(P<0.05),细胞增殖能力明显降低(P<0.05)。与PBS对照组相比,敲低SET8可抑制细胞增殖(P<0.05),而在此基础上加入LukS-PV处理后,细胞增殖抑制更加明显(P<0.05)。结论下调SET8抑制AML细胞THP-1和HL-60的增殖;且LukS-PV可以通过下调SET8抑制细胞增殖,发挥抗白血病作用。

关键词: LukS-PV, SET8, 白血病, 增殖

Abstract: Objective To explore the effect of LukS-PV on the proliferation in acute myeloid leukemia (AML) cells THP-1 and HL-60 by down-regulating methyltransferase SET8.Methods Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot were used to detect the expression levels of SET8 mRNA and protein in THP-1 and HL-60 cells after treatment with different concentrations of LukS-PV.The lentiviral vector with knock-down SET8 expression was transfected into THP-1 and HL-60 cells.SET8 knockdown efficiency was detected by qRT-PCR and Western blot,and the effects of SET8 knock-down on cell proliferation was detected by EdU assay.Finally,on the basis of knocking down SET8,adding LukS-PV treatment for detecting cell proliferation by EdU to explore whether LukS-PV inhibits the proliferation of leukemic cells THP-1 and HL-60 by down-regulating SET8.Results In AML cell lines THP-1 and HL-60,the expression of SET8 and H4K20me1 in LukS-PV treated group was significantly lower than that in PBS control group in a concentration-dependent manner (P<0.05).Compared with the negative control group,SET8 expression level was significantly decreased in THP-1 and HL-60 cells transfected with knock-down SET8 lentiviral (P<0.05),and cell proliferation ability showed the same trend (P<0.05).Compared with the PBS control group,knocking down SET8 could inhibit cell proliferation (P<0.05),and the inhibition of cell proliferation was more obvious when LukS-PV was added on this basis (P<0.05).Conclusion Down-regulating SET8 inhibits the proliferation of THP-1 and HL-60 AML cells,and LukS-PV can inhibit cell proliferation by down-regulating SET8 and play an anti-leukemia effect.

Key words: LukS-PV, SET8, Leukemia, Proliferation

中图分类号:

R551.3

施岚, 许良飞, 聂正超, 常文娇, 马筱玲. 金葡菌杀白细胞素S组分LukS-PV抑制急性髓系白血病细胞增殖的机制研究^*[J]. 临床输血与检验, 2020, 22(6): 603-609.

SHI Lan, XU Liang-fei, NIE Zheng-chao, et al. Study on the Inhibitory Mechanism of Staphylococcus Aureus Leukocidin S Component LukS-PV on the Proliferation in Acute Myeloid Leukemia Cells[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2020, 22(6): 603-609.

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金葡菌杀白细胞素S组分LukS-PV抑制急性髓系白血病细胞增殖的机制研究^*

Study on the Inhibitory Mechanism of Staphylococcus Aureus Leukocidin S Component LukS-PV on the Proliferation in Acute Myeloid Leukemia Cells

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